Journal
BIOPHYSICAL JOURNAL
Volume 96, Issue 2, Pages 707-716Publisher
CELL PRESS
DOI: 10.1016/j.bpj.2008.09.051
Keywords
-
Categories
Funding
- Cell Migration Consortium [U54 GM064346]
- National Institutes of Health [P41-RRO3155, P50-GM076516]
- Natural Sciences and Engineering Research Council of Canada
- Canadian Institutes of Health Research
Ask authors/readers for more resources
We describe a general method for detecting molecular complexes based on the analysis of single molecule fluorescence fluctuations from laser scanning confocal images. The method detects and quantifies complexes of two different fluorescent proteins noninvasively in living cells. Because in a raster scanned image successive pixels are measured at different times, the spatial correlation of the image contains information about dynamic processes occurring over a large time range, from the microseconds to seconds. The correlation of intensity fluctuations measured simultaneously in two channels detects protein complexes that carry two molecules of different colors. This information is obtained from the entire image. A map of the spatial distribution of protein complexes in the cell and their diffusion and/or binding properties can be constructed. Using this cross correlation raster image spectroscopy method, specific locations in the cell can be visualized where dynamics of binding and unbinding of fluorescent protein complexes occur. This fluctuation imaging method can be applied to commercial laser scanning microscopes thereby making it accessible to a large community of scientists.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available